期刊
VETERINARY PARASITOLOGY
卷 182, 期 2-4, 页码 287-296出版社
ELSEVIER
DOI: 10.1016/j.vetpar.2011.05.053
关键词
Haemaphysalis longicornis; Acid phosphatase; Theileria sergenti; Characterization; Anti-tick vaccine
资金
- Gansu Provincial Natural Science Foundation [096RJZA128]
- Hi-Tech Research and Development Program of China [2009AA10Z402]
The full-length cDNA encoding acid phosphatase (HL-3) from Haemaphysalis longicornis was obtained by 5' rapid amplification of cDNA ends (RACE). The cDNA contained a 1137 bp open reading frame (ORF) coding for 356 amino acids with a predicted theoretical iso-electric point (pl) of 6.35 and molecular weight of 41.0 kDa. The recombinant protein was expressed in Escherichia coli. The enzyme could hydrolyze para-nitrophenyl phosphate (pNPP) substrate at an optimum pH of 5.0. Real-time RT-PCR analysis showed that the HL-3 transcripts were expressed in various stages of unfed ticks and were significantly induced by blood feeding. Furthermore, the expression of HL-3 in midguts was significantly higher than in other tested tissues of partially fed adult ticks. The transcripts of the HL-3 mRNA in lipopolysaccharide (LPS)-injected ticks were 1.75 times of the PBS-injected control; Theileria sergenti infected larvae expressed 3.86 more times than that of uninfected ones. Western blot analysis showed that rabbit antiserum against the recombinant rHL-3 could recognize a native protein of approximately 41.0 kDa in the lysates from different stages of ticks. Vaccination of rabbits with the rHL-3 conferred partial protective immunity against ticks, resulting in 28% mortality and 10.6% reduction in engorgement weight of adult ticks, respectively. These results suggested that the HL-3 was involved in tick innate immunity and could be used as a potential candidate antigen for the development of anti-tick vaccines. Crown Copyright (C) 2011 Published by Elsevier B.V. All rights reserved.
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